Crop Protection最新文献

{"title":"First report of Stemphylium xanthosomatis causing leaf spot disease in Polygonatum cyrtonema Hua in China","authors":"Wensong Zhang ,&nbsp;Kaiyun Jiang ,&nbsp;Shuang Liang ,&nbsp;Xiangsheng Chen ,&nbsp;Zhong Li ,&nbsp;Fengquan Liu","doi":"10.1016/j.cropro.2025.107516","DOIUrl":"10.1016/j.cropro.2025.107516","url":null,"abstract":"<div><div><em>Polygonatum cyrtonema</em> Hua (“yellow essence”) belongs to the family Liliaceae and is a valuable component of Chinese medicine. In October 2024, a survey of a Chinese herbal medicine planting base in Kaili City, Guizhou Province, found that <em>P. cyrtonema</em> Hua was affected by a serious disease with an incidence of 35–60 % that substantially reduced its yield and quality and, commensurately, economic return. Koch's postulates, molecular phylogenetic analysis, and morphology confirmed <em>Stemphylium xanthosomati</em>s as the causative pathogen. This is the first report of <em>S. xanthosomatis</em> infecting <em>P. cyrtonema</em> Hua. This study enriches the pathogen diversity database of <em>P. cyrtonema</em> Hua and provides an essential theoretical foundation for disease diagnosis and targeted management strategies.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107516"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145689944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection and genome characterization of beet mosaic virus naturally infecting fodder beet crop in Argentina","authors":"Luciani Cecilia Elizabeth ,&nbsp;Celli Marcos Giovani ,&nbsp;Brugo Carivali María Florencia ,&nbsp;Bongiorno Vanina Aylen ,&nbsp;Conci Luis Rogelio ,&nbsp;Fernandez Franco Daniel ,&nbsp;Perotto María Cecilia","doi":"10.1016/j.cropro.2025.107529","DOIUrl":"10.1016/j.cropro.2025.107529","url":null,"abstract":"<div><div>Fodder beet (<em>Beta vulgaris</em> subsp. <em>vulgaris</em>) is cultivated due its tolerance to low temperatures and poor soils as it improves the structure of forage production systems. Viral diseases represent a significant threat to beet production. <em>Beet mosaic virus</em> (BtMV) is the only confirmed potyvirus infecting beets. Between 2023 and 2024, plants showing virus-like symptoms from Argentinian main production area were surveyed. Transmission electron microscopy revealed flexuous filamentous particles consistent with BtMV. Oxford Nanopore Technologies sequencing of one sample yielded the complete BtMV genome (B12AR; 9451 nt, excluding the poly (A) tail), sharing 97.47 % amino acid identity with the BtMV-Wa isolate from the USA. Additionally, sequences of Beet cryptic virus 2, a <em>Deltapartitivirus</em>, were detected. RT-PCR confirmed BtMV infection in four of seven samples collected. These results would indicate the presence of other viruses not identified in this study. Phylogenetic analysis grouped B12AR within a clade with isolates from USA and China, distinct from a divergent lineage infecting wild beets. A recombination event involving P1 and HC-Pro regions, with parental sequences from the USA and South Africa was detected. Further studies are necessary to determine the etiology of fodder beet symptoms. This is the first report and complete genome sequence of BtMV infecting fodder beet in Argentina.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107529"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145823002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"First report of Alternaria alternata causing soft rot in pitaya (Hylocereus polyrhizus) fruits in Rio Grande do Sul, Brazil","authors":"Igor Henrique de Lima Costa ,&nbsp;Gloria Emperatriz Suqui Carchi ,&nbsp;Isabela Schneid Kroning ,&nbsp;Maria Fernanda Fernandes Siqueira ,&nbsp;Frederico Schmitt Kremer ,&nbsp;Wladimir Padilha da Silva ,&nbsp;Cândida Renata Jacobsen de Farias ,&nbsp;Alvaro Renato Guerra Dias","doi":"10.1016/j.cropro.2026.107554","DOIUrl":"10.1016/j.cropro.2026.107554","url":null,"abstract":"<div><div>This study reports the first case of <em>Alternaria alternata</em> causing soft rot in pitaya (<em>Hylocereus polyrhizus</em>) fruits in Rio Grande do Sul, Brazil. Symptomatic fruits were collected from a commercial orchard, and the pathogen was isolated, characterized both morphologically and molecularly (using ITS, GAPDH, and TEF-1α <em>loci</em>), and tested for pathogenicity. The colonies exhibited mycelial growth of 5.24 ± 0.35 cm after 7 days and 8.08 ± 0.37 cm after 14 days, with olive gray centers and light brown edges. The obclavate or obpyriform conidia had transverse (1–6) and longitudinal (0–3) septa, with a short apical tip (5–22.5 μm). Molecular analysis revealed 100 % similarity with <em>A. alternata</em> sequences in GenBank, confirming the pathogen via phylogenetic analysis. Pathogenicity tests produced dark brown, depressed lesions with a clear halo, characteristic of the disease, confirming Koch's postulates. The 100 % incidence in commercial fruit crops resulted in significant economic losses, exacerbated by favorable weather conditions. Thus, this study presents the first evidence of <em>A. alternata</em> associated with post-harvest soft rot in pitaya fruits in Brazil, underscoring the need for management strategies to reduce waste and economic losses along the production chain.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107554"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146071655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterisation of pyridaben resistance in Tetranychus urticae in the Mediterranean basin","authors":"Animesha Rath ,&nbsp;Preetom Regon ,&nbsp;Svetlana Konzedalov,&nbsp;Adi Kliot","doi":"10.1016/j.cropro.2026.107552","DOIUrl":"10.1016/j.cropro.2026.107552","url":null,"abstract":"<div><div><em>Tetranychus urticae</em>, the Two-Spotted Spider Mite (TSSM) is a plant pest causing serious economic damage in various crops worldwide. The main means of controlling TSSM in the field are based on chemical pesticides but cases of reported resistance are numerous. Pyridaben is one of the pesticides used against TSSM, however reports from growers in the area indicated that it is losing its efficiency. In this study, we performed a survey testing field populations for their susceptibility to pyridaben over the years 2023–2025. We then randomly selected five populations of TSSM from this survey originating from different geographic locations and with variable resistance abilities to pyridaben. We tested these populations both for cytochrome P450-based metabolic breakdown of pyridaben and for target-site mutations. Our survey showed that pyridaben resistance is highly common in the field with 32 populations of 37 tested being resistant to a dose of pyridaben which is double the recommended dose for commercial use in the field. Furthermore, we show that resistance in most populations is likely to be metabolic, and we use a P450-inhibitor to successfully re-introduce susceptibility to three of four resistance populations. Our data sheds light on the current situation of TSSM resistance to pyridaben in the Mediterranean basin and strongly supports reduction in use of this compound in the near future.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107552"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146089437","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Light spectra and photoperiods interactively influence growth, development, and reproductive traits of Ostrinia furnacalis (Lepidoptera: Crambidae)","authors":"Yuhua Dong ,&nbsp;Sohail Abbas ,&nbsp;Aleena Alam ,&nbsp;Bilal Ahmad ,&nbsp;Muneer Abbas ,&nbsp;Khalid Ali Khan ,&nbsp;Hamed A. Ghramh ,&nbsp;Gong Xuesong ,&nbsp;Jamin Ali ,&nbsp;Xiao Han ,&nbsp;Li Qiyun ,&nbsp;Ri-Zhao Chen","doi":"10.1016/j.cropro.2026.107537","DOIUrl":"10.1016/j.cropro.2026.107537","url":null,"abstract":"<div><div>Light spectrum and photoperiod are fundamental environmental cues that regulate insect development, survival, and reproductive performance. However, despite extensive work on photoperiodic regulation of diapause in insects, the combined roles of spectral composition and photoperiod in shaping growth, development, and reproductive traits remain poorly understood in <em>Ostrinia furnacalis</em> (Guenée). In this study, we investigated the independent and interactive effects of five light spectra (blue, purple, green, red, and white) and four photoperiod regimes (LD: 14:10, 10:14, 16:08, and 08:16 h) on the growth, development, and reproductive traits of <em>O. furnacalis</em>. Using controlled laboratory experiments, we quantified key parameters, including larval survival across instars, larval and pupal durations, pupal weight, pupation rate, adult emergence rate, adult survival, and reproductive output. Both spectral quality and photoperiod significantly affected most traits, often interacting to produce spectrum-dependent photoperiodic responses. Short-wavelength light (blue, purple) generally promoted high larval survival and stable development across photoperiods. Green light extended larval and pupal development but enhanced pupal mass and fecundity. Red and white light produced variable effects across traits. These findings suggest that short wavelengths can diminish photoperiod sensitivity, whereas longer wavelengths enhance it. Our results highlight the potential role of spectral composition in shaping pest phenology and reproductive performance, providing a basis for predictive models and the development of sustainable light-based management strategies for <em>O. furnacalis</em>.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107537"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145956698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dalbulus maidis (Hemiptera: Cicadellidae) spatio-temporal dynamics and infection prevalence with the corn stunt mollicutes and virus in Santa Catarina, Brazil","authors":"Jacson Ferreira ,&nbsp;Marcos Vinicius Silva de Andrade ,&nbsp;Eduardo Silva Gorayeb ,&nbsp;Lilian Katiany Castello Rabello Zinger ,&nbsp;Daian Marcos Savaris ,&nbsp;Monalisa Cristina de Cól ,&nbsp;Emerson Medeiros Del Ponte ,&nbsp;Fábio Nascimento da Silva ,&nbsp;Leandro do Prado Ribeiro ,&nbsp;Maria Cristina Canale","doi":"10.1016/j.cropro.2025.107510","DOIUrl":"10.1016/j.cropro.2025.107510","url":null,"abstract":"<div><div>Regular and large-scale outbreaks of the diseases of the corn stunt and virus complex have been reported in Brazil since 2015, including Santa Catarina (SC) State. The pathogens are vectored by the corn leafhopper <em>Dalbulus maidis</em> (Hemiptera: Cicadellidae) and the resulting epidemics result in substantial impacts on maize production. In this study, it is reported the abundance of <em>D. maidis</em> and its infection with corn stunt mollicutes [maize bushy stunt phytoplasma (MBSP) and corn stunt spiroplasma (CSS)], and maize rayado fino virus (MRFV) in Santa Catarina during the 2021/22 and 2022/23 growing seasons. Monitoring was conducted from July to April (40 weeks) comprising both early and late planting seasons in various maize fields across the State. Each week, insect traps were inspected and brought to the laboratory where leafhoppers were counted, and molecular detection was performed for MBSP and CSS, as well as MRFV. Approximately 3000 collections were made during each growing season. Corn leafhopper counting varied in 37 % between the two seasons, being generally lower during the vegetative stage, with a marked increase during the reproductive phase. CSS was found to be more prevalent than MBSP, and MRFV was more prevalent than either of those mollicutes. Infection <em>of D. maidis</em> with MBSP, CSS, and MRFV fluctuated throughout maize development and across the forty weeks of monitoring. Moreover, the MBSP detection in corn leafhopper was inversed related with altitude. These results can serve as a decision-support tool within a translational epidemiology framework, enabling practical, field-level interventions to prevent or mitigate yield losses in maize production systems.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107510"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145614011","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The mite, the bug, and the nematode: Assessing the efficacy of three biological control agents against Echinothrips americanus (Thysanoptera: Thripidae)","authors":"Fabio Marino ,&nbsp;Linda Abenaim ,&nbsp;Giuseppe Conte ,&nbsp;Daniele Marraccini ,&nbsp;Priscilla Farina ,&nbsp;Barbara Conti","doi":"10.1016/j.cropro.2026.107553","DOIUrl":"10.1016/j.cropro.2026.107553","url":null,"abstract":"<div><div>The poinsettia thrips, <em>Echinothrips americanus</em> Morgan, 1313, is an invasive pest native to North America. The species attacks various protected ornamental and crop plants causing extensive foliage damage. Severe infestations have recently occurred in greenhouses specialized in <em>Hibiscus</em> spp. cultivation in Central Italy. Given the currently limited possibility for its integrated management, we evaluated the efficacy of three commercial biological control agents: the mite <em>Neoseiulus californicus</em> (McGregor, 1954), the bug <em>Orius laevigatus</em> (Fieber, 1860)<em>,</em> and the nematode <em>Steinernema feltiae</em> (Filipjev, 1934). Our trials were conducted in Huffaker cages under laboratory conditions, using <em>E. americanus</em> second-instar larvae as prey/host. For the first two biological control agents, we assessed predation efficiency and predation latency times. For the nematode, we determined the parasitization efficacy and optimal treatment concentration. For <em>N. californicus</em>, we observed a predation rate of 50.52% after 48 h and predation activity beginning after a mean of 3 h and 53 min. <em>Orius laevigatus</em> provided a predation rate of 73.75% after 12 h and started preying after a mean time of 7 min and 31 s. The nematode, at the tested concentrations of 50, 100, and 150 infective juveniles cm⁻² of leaf, showed a parasitization rate after 72 h of 46.59, 79.97, and 93.32%, respectively. In conclusion, <em>O. laevigatus</em> and <em>S. feltiae</em> are more effective as biological control agents against <em>E. americanus</em>. Given their different biology and behaviour, the integrated adoption of both agents for inundative release could be a valid solution for managing <em>E. americanus</em> in greenhouse hibiscus cultivation.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107553"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146071656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Flucarbazone-sodium resistance detection in Japanese brome (Bromus japonicus Thunb. ex Murr.) and candidate herbicides for its chemical control in the wheat fields of the North China Plain","authors":"Hengzhi Wang ,&nbsp;Yunxia Duan ,&nbsp;Cheng Yang ,&nbsp;Yi Li ,&nbsp;Mengmeng Hu ,&nbsp;Xinyu Sun ,&nbsp;Ping Yu ,&nbsp;Tao Jin ,&nbsp;Jinxin Wang ,&nbsp;Weitang Liu","doi":"10.1016/j.cropro.2025.107524","DOIUrl":"10.1016/j.cropro.2025.107524","url":null,"abstract":"<div><div>Japanese brome (<em>Bromus japonicus</em>), a competitive grass weed with extensive distribution in winter wheat (<em>Triticum aestivum</em>) fields across the North China Plain, is naturally tolerant to many commercial post-emergence (POST) herbicides used in wheat. Flucarbazone-sodium, an acetolactate synthase (ALS) inhibitor, is vital for the effective management of <em>B. japonicus</em> in winter wheat fields. However, resistance evolving in <em>B. japonicus</em> presents a major obstacle for maintaining sustainable weed management in winter wheat systems, and the current status of resistance remains unclear. Thus, we assessed the sensitivity of 33 <em>B. japonicus</em> populations, gathered from winter wheat fields across the North China Plain, to flucarbazone-sodium, and target-site resistance (TSR) mechanism was investigated. Moreover, the <em>B. japonicus</em> sensitivity to 20 other herbicides/herbicide mixtures was also evaluated to identify candidate herbicides for its control. The results of single-dose screen test showed that 29 populations evolved resistance (RRR and RR), 2 populations showed potential for developing resistance (R?) and 1 population was susceptible. Based on whole-plant dose-response assays, the resistant biotypes exhibited a 2.2–56.4-fold increase in resistance level to flucarbazone-sodium compared to the susceptible standard. The TSR mechanism investigation indicated that at least the decreased ALS sensitivity to flucarbazone-sodium caused by P197S and D376E mutations of ALS involved in the resistance in <em>B. japonicus</em> populations. The results of effective herbicide screening showed that POST-applied herbicide used in this study possessed limited control efficacy, with over 70 % dry weight inhibition of mesosulfuron-methyl, pyroxsulam, flucarbazone-Na + isoproturon, flucarbazone-Na + mesosulfuron-methyl and pyroxsulam + isoproturon for partial <em>B. japonicus</em> populations. Fortunately, 9 herbicides/herbicide mixtures applied pre-emergence (PRE) including acetochlor, flufenacet + diflufenican + flurtamon, pyroxasulfone, flufenacet, isoproturon + pretilachlor + halosulfuron-methyl, trifludimoxazin, cinmethylin, flufenacet + diflufenican + isoproturon, and flufenacet + isoproturon were obtained, possessing exceeding 80 % control efficacy against resistant <em>B. japonicus</em> in this study. These findings will be helpful for understanding the herbicide resistance in <em>B. japonicus</em> and its management in agricultural practice.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107524"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145784824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Block-stumping as an effective and cost-efficient style of pruning to reduce coffee berry borer (Hypothenemus hampei Ferrari) infestation and damage in Hawaiian coffee farms","authors":"Luis F. Aristizábal ,&nbsp;Melissa A. Johnson ,&nbsp;Mark G. Wright","doi":"10.1016/j.cropro.2025.107502","DOIUrl":"10.1016/j.cropro.2025.107502","url":null,"abstract":"<div><div>Pruning is a cultural practice conducted by coffee farmers to increase production and facilitate the management of insect pests and diseases. This field study assessed the impact of different pruning styles on the regulation of coffee berry borer (CBB, <em>Hypothenemus hampei</em>), widely considered the most damaging insect pest of coffee worldwide. For each pruning system (block-stumping, Kona style, topping, and Beaumont-Fukunaga), 3–4 coffee lots were selected on Hawaiʻi Island and CBB infestation, bean damage, and harvesting efficacy was evaluated in 2023 and 2024. Yields, sale price, and pruning costs were also recorded to calculate the benefit cost ratio of each pruning style. Block-stumping resulted in significantly lower CBB infestation (0.6 % and 2.6 %, year one and two of harvest) relative to Beaumont-Fukunaga (6.3 %), topping (8.6 %) and Kona style (10.0 %). Block-stumping also resulted in significantly lower bean damage (2.6 %) compared to topping (24.3 %) and Kona style (25.6 %). The most effective harvesting was done in lots that were block-stumped (≤6.7 berries left per tree), while Beaumont-Fukunaga (10.9), topping (14.5), and Kona style (15.7) resulted in low harvesting efficacy. While there was no significant difference among pruning styles, the benefit cost ratio was higher in lots that employed block-stumping (2.29) or Beaumont-Fukunaga (2.29) compared to topping (2.18) and the traditional Kona style (2.04). Our findings suggest that block-stumping is the optimal pruning style for reducing CBB infestation and bean damage, and results in improved harvesting efficacy and a higher benefit cost ratio relative to other pruning styles that are commonly used in Hawaiʻi.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107502"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145619681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Crude sap-based CRISPR-Cas12a-assisted RT-RPA assay for specific detection of citrus yellow vein clearing virus in citrus","authors":"Rakesh Kumar ,&nbsp;Susheel Kumar Sharma ,&nbsp;Nishant Srivastava ,&nbsp;Pooja Bhardwaj ,&nbsp;Sini Kumari ,&nbsp;Abhinav Rawat ,&nbsp;Viswanathan Chinnusamy ,&nbsp;Virendra Kumar Baranwal ,&nbsp;Nitika Gupta","doi":"10.1016/j.cropro.2026.107540","DOIUrl":"10.1016/j.cropro.2026.107540","url":null,"abstract":"<div><div>Citrus yellow vein clearing virus (CYVCV) is an emerging mandarivirus globally, commonly present in citrus orchards in India. Early detection of CYVCV is essential to prevent its spread through propagative material and to support integrated disease management. Conventional RT-PCR, although reliable and widely used, are time-intensive, require RNA extraction, and depend on sophisticated laboratory infrastructure, which limits their application for rapid on-site surveillance and large-scale indexing. In this study, we developed and validated a CRISPR-Cas12a-assisted reverse transcription recombinase polymerase amplification (RT-RPA) assay using crude-sap as template for rapid, sensitive, and sequence-specific detection of CYVCV. A 241 bp RT-RPA amplicon derived from the RNA-dependent RNA polymerase (RdRp) region of CYVCV was specifically recognized by a Cas12a-crRNA-complex, which activated collateral cleavage of a fluorescent single-stranded DNA-reporter, generating robust and unambiguous signals within 40–50 min. The analytical sensitivity of the CRISPR-based system was established at the 10⁻⁶ dilution of RT-RPA product by quantitative fluorescence measurement and was further translated into absolute copy numbers using a plasmid-based SYBR Green qPCR standard curve, while endpoint visualization using a UV transilluminator was feasible up to the 10⁻³ dilution. Validation on symptomatic citrus field samples revealed strong fluorescence signals in infected plants (up-to 91,447 AU), whereas healthy and non-template controls consistently remained at baseline. Importantly, visible fluorescence in infected samples under UV light further underscored the field-deployable potential of the assay. Compared with conventional RT-PCR, the CRISPR-based RT-RPA platform demonstrated an approximately ∼1.4-fold per-sample cost reduction, supported by itemized reagent-level cost analysis. These attributes establish the assay as robust, user-friendly, and scalable diagnostic-tool for CYVCV detection, offering strong potential for nursery indexing, orchard surveillance, and certification programs.</div></div>","PeriodicalId":10785,"journal":{"name":"Crop Protection","volume":"202 ","pages":"Article 107540"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145995179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}